Immunogenic compositions for herpes simplex virus proteins

Abstract

The disclosure provides live attenuated measles vectors encoding in their genome one or more heterologous genes for herpes simplex virus (HSV) proteins, e.g., glycoprotein B, glycoprotein C, glycoprotein D, and variants thereof, as well as nucleic acid constructs encoding such measles vectors. The disclosure also relates to immunogenic compositions comprising live attenuated measles vectors encoding HSV proteins, immunogenic compositions comprising such measles vectors, and use of such measles vectors and immunogenic compositions to induce an immune response to HSV in subjects.

1 . An isolated nucleic acid molecule comprising:
 a) a cDNA encoding a full length, antigenomic (+) RNA strand of an attenuated strain of measles virus (MV-cDNA);   b) one or more cDNAs encoding an herpes simplex virus (HSV) protein (HSV cDNA) selected from the group consisting of: gC, gD, gB, UL19, and variants thereof;   c) an upstream additional transcriptional unit (ATU) cDNA operably linked to the HSV cDNA that is 5′ of the HSV cDNA (upstream ATU cDNA); and   d) a downstream ATU cDNA operably linked to the HSV cDNA that is 3′ of the HSV cDNA (downstream ATU cDNA);   wherein the upstream ATU cDNA, the HSV protein cDNA, and the downstream ATU cDNA are between P and M genes of the MV-cDNA at ATU2 or between H and L genes of the MV-cDNA at ATU3.   
     
     
         2 . The isolated nucleic acid molecule of  claim 1 , wherein each of the one or more HSV cDNAs encodes an HSV protein sequence independently selected from the group consisting of: SEQ ID NO: 54 (HSV-2 gC, wild-type); SEQ ID NO: 57 (HSV-2 gC F327A);
 SEQ ID NO: 55 (HSV-2 gB, wild-type); SEQ ID NO: 58 (HSV-2 gBmut); SEQ ID NO: 58.1 (HSV-2 gBmutdel25); SEQ ID NO: 64 (HSV-2 gBwtdel25); SEQ ID NO: 56 (HSV-2 gD, wild-type); and SEQ ID NO: 65 (HSV-2 UL19).   
     
     
         3 . The isolated nucleic acid of  claim 1 , wherein the upstream ATU cDNA, the HSV cDNA, and the downstream ATU cDNA is at ATU2 or ATU3 in the MV-cDNA. 
     
     
         4 . (canceled) 
     
     
         5 . The isolated nucleic acid molecule of  claim 1 , wherein the upstream ATU cDNA sequence is set forth in SEQ ID NO: 87 or 90. 
     
     
         6 . (canceled) 
     
     
         7 . The isolated nucleic acid molecule of  claim 1 , comprising a sequence selected from the group consisting of SEQ ID NOs: 59-63 and 96-97. 
     
     
         8 . An isolated nucleic acid molecule comprising:
 a) a cDNA encoding a full length, antigenomic (+) RNA strand of an attenuated strain of measles virus (MV-cDNA);   b) a first cDNA encoding an herpes simplex virus (HSV) protein selected from the group consisting of: gC, D, gB, UL19, and variants thereof (first HSV cDNA);   c) a second HSV cDNA encoding an HSV protein selected from the group consisting of: gC, gD, gB, UL19, and variants thereof (second HSV cDNA), wherein the first and second HSV cDNAs do not have the same sequence;   d) an upstream additional transcriptional unit (ATU) cDNA that is 5′ of the first HSV cDNA (upstream ATU cDNA);   e) a downstream ATU cDNA that is 3′ of the second HSV cDNA (downstream ATU cDNA); and   f) an interstitial ATU cDNA between the first and second HSV cDNAs (interstitial ATU cDNA);   wherein the upstream ATU cDNA, the first and second HSV cDNAs, the interstitial ATU cDNA and the downstream ATU cDNA are operably linked; and   wherein the upstream ATU cDNA, the first and second HSV cDNAs, the interstitial ATU, and the downstream ATU cDNA are between P and M genes of the MV-cDNA at ATU2 or between H and L genes of the MV-cDNA at ATU3.   
     
     
         9 . The isolated nucleic acid molecule of  claim 8 , wherein the first and second HSV cDNAs each encode an HSV protein sequence independently selected from the group consisting of SEQ ID NO: 54 (HSV-2 gC, wild-type); SEQ ID NO: 57 (HSV-2 gC F327A); SEQ ID NO: 55 (HSV-2 gB, wild-type); SEQ ID NO: 58 (HSV-2 gBmut); SEQ ID NO: 58.1 (HSV-2 gBmutdel25); SEQ ID NO: 64 (HSV-2 gBwtdel25); SEQ ID NO: 56 (HSV-2 gD, wild-type);
 and SEQ ID NO: 65 (HSV-2 UL19).   
     
     
         10 . The isolated nucleic acid of  claim 8 , wherein the upstream ATU cDNA, the first and second HSV cDNAs, and the downstream ATU cDNA are at ATU2 or ATI3 in the MV-cDNA. 
     
     
         11 . (canceled) 
     
     
         12 . The isolated nucleic acid molecule of  claim 8 , wherein the upstream ATU cDNA sequence is set forth in SEQ ID NO: 87 or 90. 
     
     
         13 . (canceled) 
     
     
         14 . The isolated nucleic acid molecule of  claim 8 , wherein the interstitial ATU cDNA sequence is selected from the group consisting of SEQ ID NOs: 83, 87, 90, and 92. 
     
     
         15 . The isolated nucleic acid molecule of  claim 8 , comprising a sequence selected from the group consisting of SEQ ID NOs: 66, 68, 69, and 70. 
     
     
         16 . An isolated nucleic acid molecule comprising:
 a) a cDNA encoding a full length, antigenomic (+) RNA strand of an attenuated strain of measles virus (MV-cDNA);   b) a first cDNA encoding an herpes simplex virus (HSV) protein (HSV cDNA) selected from the group consisting of: gC, gD, gB, UL19, and variants thereof (first HSV cDNA);   c) a second HSV cDNA encoding an HSV protein selected from the group consisting of: gC, gD, gB, UL19, and variants thereof (second HSV cDNA), wherein the first and second HSV cDNAs do not have the same sequence;   d) an upstream additional transcriptional unit (ATU) cDNA that is 5′ of the first HSV cDNA (upstream ATU cDNA);   e) a downstream ATU cDNA that is 3′ of the second HSV cDNA (downstream ATU cDNA); and   f) a furin cleavage site cDNA and 2A peptide cDNA between the first and second HSV protein cDNAs (Fur-2A cDNA);   wherein the upstream ATU cDNA, the first and second HSV cDNAs, the Fur-2A cDNA, and the downstream ATU cDNA are operably linked; and   wherein the upstream ATU cDNA, the first and second HSV cDNAs, the Fur-2A cDNA, and the downstream ATU cDNA are between P and M genes of the MV-cDNA at ATU2 or between H and L genes of the MV-cDNA at ATU3.   
     
     
         17 . The isolated nucleic acid molecule of  claim 16 , wherein the first and second HSV cDNAs each encode an HSV protein sequence independently selected from the group consisting of: SEQ ID NO: 54 (HSV-2 gC, wild-type); SEQ ID NO: 57 (HSV-2 gC F327A); SEQ ID NO: 55 (HSV-2 gB, wild-type); SEQ ID NO: 58 (HSV-2 gBmut); SEQ ID NO: 
     
     
       58. 1 (HSV-2 gBmutdel25); SEQ ID NO: 64 (HSV-2 gBwtdel25); SEQ ID NO: 56 (HSV-2 gD, wild-type); and SEQ ID NO: 65 (HSV-2 UL19). 
     
     
         18 . The isolated nucleic acid of  claim 16 , wherein the upstream ATU cDNA, the first and second HSV cDNAs, the Fur-2A cDNA, and the downstream ATU cDNA are at ATU2 or ATU3 in the MV-cDNA. 
     
     
         19 . (canceled) 
     
     
         20 . The isolated nucleic acid molecule of  claim 16 , wherein the upstream ATU cDNA sequence is set forth in SEQ ID NO: 87 or 90. 
     
     
         21 . The isolated nucleic acid molecule of  claim 16 , wherein the downstream ATU cDNA sequence is set forth in SEQ ID NO: 90. 
     
     
         22 . The isolated nucleic acid molecule of  claim 16 , wherein the furin cDNA of the Fur-2A cDNA encodes a protein sequence selected from the group consisting of SEQ ID NOs: 14-53, and wherein the 2A peptide cDNA of the Fur-2A cDNA encodes a protein sequence selected from the group consisting of SEQ ID NOs: 4-11. 
     
     
         23 - 49 . (canceled) 
     
     
         50 . A vector for the rescue of a recombinant measles virus, comprising the isolated nucleic acid molecule of  claim 1 . 
     
     
         51 - 54 . (canceled) 
     
     
         55 . A recombinant measles virus comprising in its genome a cDNA sequence comprising the nucleic acid molecule of  claim 1 . 
     
     
         56 . An immunogenic composition comprising (i) an effective amount of the recombinant measles virus of  claim 55 , and (ii) a pharmaceutically acceptable carrier. 
     
     
         57 . A method for treating or preventing a herpes simplex virus (HSV) infection in a subject in need thereof, comprising administering an effective amount of the immunogenic composition according to  claim 56  to the subject. 
     
     
         58 . A method for inducing a protective immune response against herpes simplex virus (HSV) in a subject in need thereof, comprising administering an effective amount of the immunogenic composition of  claim 56  to the subject. 
     
     
         59 - 65 . (canceled) 
     
     
         66 . An isolated peptide comprising the sequence set forth in SEQ ID NO: 58 (HSV-2 gBmut), SEQ ID NO: 58.1 (HSV-2 gBmutdel25), or SEQ ID NO: 64 (HSV-2 gBwtdel25),
 wherein SEQ ID NOs: 58.1 and 64 do not include residues 877-901 of SEQ ID NO: 55 (HSV-2 gB wild-type), or variants thereof, and   wherein SEQ ID NOs: 58 and 58.1 comprise an alanine at position 665, an alanine at position 675, and an alanine at position 677.   
     
     
         67 - 68 . (canceled) 
     
     
         69 . An isolated nucleic acid molecule encoding the isolated peptide of  claim 66 . 
     
     
         70 . An immunogenic composition comprising (i) an effective amount of the isolated peptide of  claim 66 , and (ii) a pharmaceutically acceptable carrier. 
     
     
         71 . A method for treating or preventing a herpes simplex virus (HSV) infection in a subject in need thereof, comprising administering an effective amount of the immunogenic composition according to  claim 70  to the subject. 
     
     
         72 . A method for inducing a protective immune response against herpes simplex virus (HSV) in a subject in need thereof, comprising administering an effective amount of the immunogenic composition of  claim 70  to the subject. 
     
     
         73 - 79 . (canceled)